Retinoic acid receptor γ activation promotes differentiation of human induced pluripotent stem cells into esophageal epithelium

Abstract

Background The esophagus is known to be derived from the foregut. However, the mechanisms regulating this process remain unclear. In particular, the details of the human esophagus itself have been poorly researched. In this decade, studies using human induced pluripotent stem cells (hiPSCs) have proven powerful tools for clarifying the developmental biology of various human organs. Several studies using hiPSCs have demonstrated that retinoic acid (RA) signaling promotes the differentiation of foregut into tissues such as lung and pancreas. However, the effect of RA signaling on the differentiation of foregut into esophagus remains unclear. Methods We established a novel stepwise protocol with transwell culture and an air–liquid interface system for esophageal epithelial cell (EEC) differentiation from hiPSCs. We then evaluated the effect of all-trans retinoic acid (ATRA), which is a retinoic acid receptor (RAR)α, RARβ and RARγ agonist, on the differentiation from the hiPSC-derived foregut. Finally, to identify which RAR subtype was involved in the differentiation, we used synthetic agonists and antagonists of RARα and RARγ, which are known to be expressed in esophagus. Results We successfully generated stratified layers of cells expressing EEC marker genes that were positive for lugol staining. The enhancing effect of ATRA on EEC differentiation was clearly demonstrated with quantitative reverse transcription polymerase chain reaction, immunohistology, lugol-staining and RNA sequencing analyses. RARγ agonist and antagonist enhanced and suppressed EEC differentiation, respectively. RARα agonist had no effect on the differentiation. Conclusion We revealed that RARγ activation promotes the differentiation of hiPSCs-derived foregut into EECs.

Alisol B Alleviates Hepatocyte Lipid Accumulation and Lipotoxicity via Regulating RARα-PPARγ-CD36 Cascade and Attenuates Non-Alcoholic Steatohepatitis in Mice

“…Alisol B (purity > 97%) was extracted and purified, and the structure was confirmed by comparing the 1 H, 13 C NMR, and MS data with reported data [29]. Dried rhizomes of Alisma orientale were extracted three times with 95% ethanol and then concentrated under vacuum to yield a crude extract, which was suspended in H 2 O followed by partition with AcOEt.…”

“…Retinoic acid receptors (RARs), composed of three subtypes (RARα, RARβ, and RARγ), belong to the steroid/thyroid superfamily of nuclear hormone receptors. RARα is expressed in most tissues, whereas RARβ is mainly expressed in central nervous system [12] and RARγ is mainly expressed in skin, epithelium, and cartilage [13]. RARs are activated by the biologically active metabolite of retinol such as all-trans retinoic acid (atRA) and 9-cis retinoic acid [14], and form heterodimers with retinoid X receptor (RXR) to regulate gene transcription and signal transduction pathways.…”

Differentiation‑related zinc finger protein 750 suppresses cell growth in esophageal squamous cell carcinoma

“…The results of the present study demonstrated the involvement of Zfp750 in the esophageal keratinocyte differentiation program through the upregulation and downregulation of Zfp750 expression with chemical stimulation and inhibition, by ATRA and Compound E, respectively, of the differentiation program in mNEEOs. ATRA is a chemical promoter of keratinocyte differentiation in esophageal cells (28,29) and was used in the present study to induce the epithelial differentiation program. Alternatively, γ-secretase inhibitor Compound E was used to mimic the inhibition of the keratinocyte differentiation program through the inhibition of the NOTCH signaling pathway (30,31).…”

“…RNA was extracted as aforementioned from the ATRA-treated cells at 48 h post-treatment and subjected to RT-qPCR analysis. Experiments were performed on KYSE30 and KYSE180 cell lines using previously reported ATRA concentrations as a reference to determine the optimal ATRA concentration for each cell line (28).…”

Epithelial-derived factors induce muscularis mucosa of human induced pluripotent stem cell-derived gastric organoids

“…The scarcity of specimens of human embryo and fetal organs has hindered studies on human developmental biology. In previous decades, various organs and tissues derived from the endoderm, mesoderm, and ectoderm have become able to be induced from human pluripotent stem cells, such as human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) (Koterazawa et al, 2020;Shi et al, 2017;Suzuki et al, 2019), which has allowed us to investigate normal and pathological human developmental processes and their underlying mechanisms. McCracken et al reported the generation of human pluripotent stem cell-derived gastric organoids (hGOs); however, the hGOs lacked subepithelial SM tissue (McCracken et al, 2014).…”